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標題: Opposing action of estrogen receptors alpha and beta on tumor necrosis factor-alpha gene expression and caspase-8-mediated apoptotic effects in HA22T cells
作者: Huang, E.J.
Wu, C.C.
Lee, S.D.
Chen, J.H.
Liu, J.Y.
Ko, J.L.
Lin, J.A.
Lu, M.C.
Chen, L.M.
Huang, C.Y.
Kuo, W.W.
關鍵字: apoptotic effects;estrogen receptor-beta;HA22T cells;tumor necrosis;factor-alpha;human hepatoma-cells;breast-cancer cells;messenger-rna;er-beta;hepatocellular-carcinoma;cyclin d1;chromosomal localization;mediated;inhibition;prostate-cancer;activation
Project: Molecular and Cellular Biochemistry
期刊/報告no:: Molecular and Cellular Biochemistry, Volume 287, Issue 1-2, Page(s) 137-145.
Hepatocellular carcinoma (HCC), the major manifestation of primary liver cancer, is one of the most frequent and malignant cancers worldwide, especially in Taiwan. Estrogen receptors (ERs) have been reported to play either a proliferation- or apoptosis-enhancing role in the differentiation of cancers, including HCC. In a previous experiment, we showed that transient overexpressed estrogen receptor-alpha induced early stage HCC cell line Hep 3B cell apoptosis by increasing the hTNF-alpha gene expression in a ligand-independent manner. To further clarify if the apoptotic effect occurs in poorly differentiated HCC cell line, HA22T, and elucidate the roles of ERs and TNF-alpha, DNA fragmentation and caspase activity were measured in late stage HCC cell line, HA22T, by measuring the expression of hER-alpha and hER-beta using a Tetracycline-induciable system (Tet-on). Increased DNA fragmentation and caspase-3 activity were found in hER beta-overexpressed HA22T cells treated with estrogen (10(-8) M) but not in hER alpha-overexpressed HA22T cells. Using RT-PCR/PCR and western blotting in HA22T cells, overexpressed hER-beta was also found to increase the expression of hTNF-alpha mRNA and induce hTNF-alpha-dependent luciferase activity in a ligand-dependent manner. Additionally, LPS treatment and hER-beta overexpression both enhance caspase-8 activities, whereas neither hER-beta nor E-2 treatment affected caspase-9 activities. In addition, the overexpressed hER-beta plus E-2 enhanced DNA fragmentation and caspase-8 activities were only partially reduced by anti-hTNF-alpha (0.1ng/ml), which was possibly due to the involvement of P53 and TGF-beta. Taken together, our data indicates that overexpressed hER-beta but not hER-alpha may induce caspase-8-mediated apoptosis by increasing the hTNF-alpha gene expression in a ligand-dependent manner in poorly differentiated HA22T cells.
ISSN: 0300-8177
DOI: 10.1007/s11010-005-9092-4
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